Generation and characterization of early stage oral cancer cell line of buccal mucosa of Indian origin

Being topmost cancer in India, oral cancer management warrants discovery of novel biomarkers, treatment strategies, and targets to help with early diagnosis, treatment, and recovery. To have a continuous supply of cells, the study was aimed at generation and characterization of established cell line from buccal mucosa (BM) tumors from patients of Indian origin which can be developed as a pre-clinical tool for biomedical application. Surgically resected tumor tissue from histo-pathologically confirmed oral cancer were processed for explant culture. TBM-02 cell line was passaged and characterized for morphology and function. Further, the cell line was silenced for inflammasome pathway gene NLRP3 to evaluate its linkage with oral cancer tumorigenesis. TBM-02, successfully established from BM, was maintained up to 100 passages, exhibited epithelioid morphology, high EpCam expression and triploid ploidy with chromosomal aberrations. Novelty and human origin of TBM-02 was authenticated by Short Tandem Repeats profiling and comparison with DSMZ database. TBM-02 revealed tumorigenic potential in vitro and in vivo which was abrogated on silencing NLRP3. Increased expression of NLRP3, hallmark of chronic inflammation in TBM-02, was validated at protein and gene level and in xenograft. TBM-02 demonstrated migratory potential and was found to be a sensitive tool to study drug response. RNA sequencing demonstrated upregulation of oral cancer-associated genes and pathways. Thus, in current study, we have reported development of novel cell line from early-stage buccal mucosa cancer patient which has a strong potential to be developed and to be used as pre-clinical model for improving oral cancer management and therapeutics.